© 2020 McCann et al.Systematic genetic interaction profiles can reveal the mechanism-of-action of bioactive compounds. The imipridone ONC201, which is currently in cancer clinical trials, has been ascribed a variety of different targets. To investigate the genetic dependencies of imipridone action, we screened a genome-wide CRISPR knockout library in the presence of either ONC201 or its more potent analog ONC212. https://www.selleckchem.com/products/SB-203580.html Loss of the mitochondrial matrix protease CLPP or the mitochondrial intermediate peptidase MIPEP conferred strong resistance to both compounds. Biochemical and surrogate genetic assays showed that impridones directly activate CLPP and that MIPEP is necessary for proteolytic maturation of CLPP into a catalytically competent form. Quantitative proteomic analysis of cells treated with ONC212 revealed degradation of many mitochondrial as well as non-mitochondrial proteins. Prompted by the conservation of ClpP from bacteria to humans, we found that the imipridones also activate ClpP from Escherichia coli, B. subtilis and Staphylococcus aureus in biochemical and genetic assays. ONC212 and acyldepsipeptide (ADEP)-4, a known activator of bacterial ClpP, caused similar proteome-wide degradation profiles in S. aureus ONC212 suppressed the proliferation of a number of Gram-positive (S. aureus, B. subtilis, Enterococcus faecium) and Gram-negative species (E. coli, Neisseria gonorrhoeae). Moreover, ONC212 enhanced the ability of rifampin to eradicate antibiotic-tolerant S. aureus persister cells. These results reveal the genetic dependencies of imipridone action in human cells and identify the imipridone scaffold as a new entry point for antibiotic development. Copyright © 2020, Genetics.Normothermic perfusion provides a means to rescue steatotic liver grafts including by pharmacological defatting. In this study, we tested the potential of new drug combinations to trigger defatting in three human culture models, primary hepatocytes with induced steatosis or isolated from steatotic liver, and precision-cut liver slices (PCLS) of steatotic liver. Forskolin, L-carnitine and a PPARα agonist, all were combined with rapamycin, an immunosuppressant that induces autophagy, in a D-FAT cocktail. D-FAT was tested alone or in combination with necrosulfonamide, an inhibitor of mixed lineage kinase domain-like involved in necroptosis. Within 24 hours in all three models, D-FAT induced a decrease in triglyceride content by 30%, attributable to an up-regulation of genes involved in free fatty acid β-oxidation and autophagy, and a down-regulation of those involved in lipogenesis. Defatting was accompanied by a decrease in endoplasmic reticulum stress and in the production of reactive oxygen species. The addition of necrosulfonamide increased the efficacy of defatting by 8%-12% in PCLS, with a trend towards increased autophagy. In conclusion, culture models notably PCLS are insightful to design strategies of liver graft rescue. Defatting can be rapidly achieved by combinations of drugs targeting mitochondrial oxidative metabolism, macro-autophagy, and lipogenesis. © 2020. Published by The Company of Biologists Ltd.OBJECTIVE This study explored the changes of global public interest in internet search of ankylosing spondylitis (AS) based on Google Trends (GT) data, in order to reflect the characteristics of AS itself. METHODS GT was used to obtain the search popularity scores of the term 'AS' on a global scale, between January 2004 and December 2018, under the 'health' classification. Based on the global search data of AS provided by GT, the cosinor analysis was used to test whether there was seasonality in AS. RESULTS In general, AS related search volume demonstrated a decreasing trend from January 2004 to December 2014 and then remain stable from January 2015 to December 2018. No obvious seasonal variations were detected in AS related search volume (amplitude=1.54; phase month=3.9; low point month=9.9; p&gt;0.025), which peaked in April and bottomed out in October. The top 17 rising topics were adalimumab, spondylolisthesis, Morbus, Vladimir Mikhailovich Bekhterev, autoimmune disease, rheumatoid arthritis, ankylosis, HLA- B27 positive, Crohn's disease, rheumatology, spondylosis, arthritis, uveitis, rheumatism, sacroiliac, psoriatic arthritis and spondylitis. CONCLUSIONS Globally, there is no significant seasonal variation in GT for AS. The top fast-growing topics related to AS may be beneficial for doctors to provide targeted health education of the disease to patients and their families. © Author(s) (or their employer(s)) 2020. No commercial re-use. See rights and permissions. Published by BMJ.BACKGROUND Canadian health care facilities must report losses or thefts of opioids to Health Canada. To broaden the understanding of opioid loss in Canada, we analyzed data describing these losses to estimate the amount of opioid lost, estimate the wholesale and street value, compare the distribution of loss types between facility types and compare loss trends. METHODS We analyzed Health Canada records of losses of codeine, fentanyl, hydromorphone, morphine and oxycodone reported by Canadian facilities from January 2012 to September 2017. We conducted descriptive analyses of the opioid losses by calculating milligrams of drug lost, oral morphine equivalents, daily defined doses, approximate wholesale value and approximate street value, and compared loss trends when counted by incidents, dosage units or milligrams. RESULTS There were 64 963 reports of loss of codeine, fentanyl, hydromorphone, morphine or oxycodone over the study period. Over 112 kg of opioids were lost, an estimated $8.7 million in wholesale closses. Copyright 2020, Joule Inc. or its licensors.Carbapenem resistance in Enterobacterales is a public health threat. Klebsiella pneumoniae carbapenemase (encoded by alleles of the bla KPC family) is one of the commonest transmissible carbapenem resistance mechanisms worldwide. The dissemination of bla KPC has historically been associated with distinct K. pneumoniae lineages (clonal group 258 [lsqb]CG258[rsqb]), a particular plasmid family (pKpQIL), and a composite transposon (Tn4401). In the UK, bla KPC has represented a large-scale, persistent, management challenge for some hospitals, particularly in North-West England. The dissemination of bla KPC has evolved to be polyclonal and poly-species, but the genetic mechanisms underpinning this evolution have not been elucidated in detail; this study used short-read whole genome sequencing of 604 bla KPC-positive isolates (Illumina) and long-read assembly (PacBio)/polishing (Illumina) of 21 isolates for characterisation. We observed the dissemination of bla KPC (predominantly bla KPC-2; 573/604 [lsqb]95%[rsqb] isolates) across eight species and more than 100 known sequence types.